Vitamin D compounds and method of preparing these compounds

ABSTRACT

The invention relates to a new vitamin D compound of the general formula ##STR1## wherein: R 1  is a hydrogen atom or a hydroxy group; 
     R 2  is a (C 1  -C 3 )alkyl group, a hydroxy(C 1  -C 3 )alkyl group, a (C 1  -C 2 )alkoxymethyl group or a (C 2  -C 3 )alkenyl or alkynyl group; 
     R 3  is a branched or non-branched, saturated or unsaturated aliphatic 3- to 5 -membered hydrocarbon or oxahydrocarbon biradical, having at least 3 atoms in the main chain and being optionally substituted with one or more substituents selected from epoxy, fluoro and hydroxy; 
     R 4  is a sec. or tert. (C 3  -C 6 )alkyl group or a (C 3  -C 6 )cycloalkyl group; and 
     A and B are each individually hydrogen atoms or methyl groups, or 
     A and B form together a methylene group. 
     The invention further relates to a method of preparing these compounds and to their use in pharmacotherapy and cosmetics.

The invention relates to new vitamin D compounds, to a method ofpreparing these compounds and to their use in pharmacotherapy andcosmetics. The invention further relates to valuable new intermediates.It is generally known, that vitamin-D compounds or vitamin-D relatescompounds ("vitamin-D compounds") have a strong biological activity andmay be used in all those cases in which problems with the calciummetabolism play a part. A few years ago it was found that various activevitamin-D compounds also have other pharmacotherapeutic activities andmay be used successfully, for example, for the treatment of certain skinand bone diseases, for cosmetic applications and for treating diseaseswhich are related to cell differentiation, cell proliferation orimbalance in the immune system, including diabetes mellitus,hypertension and inflammatory diseases such as rheumatoid arthritis andasthma. In addition, these compounds may be used in various veterinaryapplications, and for diagnostic purposes.

Vitamin D compounds which are of interest for the above applications arehydroxylated vitamin D compounds, in particular vitamin D compoundshydroxylated in the 1α-, 24- and/or 25-positions. Recent developments inthe field of active vitamin D compounds are 19-nor-vitamin compounds(EP-A-038077) and C.sub.,8 -modified vitamin D compounds (EP-A-0521550),preferably also hydroxylated in the 1α-position and optionally in theC₁₇ -side chain. Other modifications of the C₁₇ -side chain have beenproposed, likewise to improve the intended activity and to suppressdetrimental side-effects. Examples of modifications of the C₁₇ -sidechain are chain elongations (homo compounds), 22-oxa modifications,fluor substitutions, epoxy groups (e.g. WO 92/21695), etc. In additioncertain 24-cyclopropyl-modified vitamin D compounds are disclosed inliterature, e.g. in WO 87/00834 (for treating abnormal celldifferentiation and proliferation) and in an article by Farach-Carson etal. in Endocrinology 1991, 129, 1876-84. Generally, however, the aboveC₁₇ -side chain modified vitamin D compounds are still not completelysatisfactory as regards their selective activity, i.e. the intendedactivity without detrimental side-effects. Further, the accessibility ofthe C₁₇ -side chain modified vitamin D compounds is often insufficientor unattractive. As an example, the preparation of the above vitamin Dcompound disclosed by Farach-Carson et al, seems very laborious, whilethe C₁₇ -side chain build-up, described in the above WO 87/00834, alsorequires various laborious synthetic steps, using a not readilyavailable ketone as a synthon. In this connection there is a need forbetter accessible C₁₇ -side chain modified vitamin D compounds. As amatter of fact, both the starting compounds for the preparation of suchvitamin-D compounds must be easily available or accessible, and themultistep preparation process must lead to the intended purpose withsufficient selectivity and efficiency.

It is therefore the objective of the present invention to provide a newclass of vitamin D compounds, which is well accessible from readilyavailable or accessible starting materials.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1 and 2 show the binding of vitamin D compounds to human vitamin Dbinding proteins, as per Example VIII.

FIGS. 3 and 4 show the differentiating effect of vitamin D compounds onhuman leukemia cells of the HL-60 line, as per Example IX.

According to the present invention this objective can be achieved with anew vitamin D compound of the general formula ##STR2## wherein: R₁ is ahydrogen atom or a hydroxy group;

R₂ is a (C₁ -C₃)alkyl group, a hydroxy(C₁ -C₃)alkyl group, a (C₁-C₂)alkoxymethyl group or a (C₂ -C₃)alkenyl or alkynyl group;

R₃ is a branched or non-branched, saturated or unsaturated aliphatic 3-to 5-membered hydrocarbon or oxahydrocarbon biradical, having at least 3atoms in the main chain and being optionally substituted with one ormore substituents selected from epoxy, fluoro and hydroxy;

R₄ is a sec. or tert. (C₃ -C₆)alkyl group or a (C₃ -C₆)cycloalkyl group;and

A and B are each individually hydrogen atoms or methyl groups, or A andB form together a methylene group.

The above new vitamin D compounds of the invention, presented by thegeneral formula I, are valuable substances. The biological results, asillustrated in the Examples, indicate that these compounds are promisingas biologically active substances and may be used in all above-mentionedpharmacotherapeutic indications, more in particular for the treatment ofosteoporosis, renal osteodystrophy, osteomalacia, skin disorders such aspsoriasis (and other hyperproliferative skin diseases), eczema anddermatitis, myopathy, leukemia, breast and colon cancer, osteosarcomas,squamous cell carcinomas, melanoma, certain immunological disorders, andtransplant rejections.

Furthermore, the new vitamin D compounds of the invention may be usedfor wound healing and may be incorporated in cosmetic compositions, suchas creams, lotions, ointments and the like, in order to preserve,condition and/or protect the skin and to improve various skinconditions, such as wrinkles, dry skin, skin slackness and insufficientsebum secretion. The new vitamin D compounds may also be used fordiagnostic purposes.

Suitable examples of the above substituent R₄ are: isopropyl,cyclopropyl, tert.-butyl, thexyl (1,1,2-trimethylpropyl), 3-pentyl andcyclopentyl.

A vitamin D compound is preferred, having the above general formula I,wherein:

R₁ is a hydroxy group;

R₂ has the meaning given above;

R₃ is a biradical of the formula --O--CH₂ --(CH₂)_(n) --, --CH₂ --CH₂--(CH₂)_(n) --, --CH═CH--(CH₂)_(n) -- or --CH₂ --CH₂ --CH(CH₃)--,

wherein n is 1 or 2;

R₄ is an isopropyl group, a cyclopropyl group or a tert.-butyl group;and A and B are hydrogen atoms or form together a methylene group.

Examples of pre-eminently suitable vitamin D compounds according to theinvention are vitamin D compounds of the above general formula I,wherein the symbols R₁, R₂, R₃, A and B have the above-defined meanings,and R₄ is an isopropyl group; because of their extremely favourablebiological properties.

It is a special merit of the present invention that the above newvitamin D compounds of the invention can easily be prepared from readilyavailable starting materials. In particular, it has been found, that thedesired C₂₅ -configuration, i.e. the attachment of the appropriatesubstituents to C₂₅, can easily be achieved by starting from a readilyaccessible ester compound.

Consequently, the invention also relates to a method of preparing avitamin D compound of the general formula I, as defined above, whereinR₁ is a hydroxy group, which method is characterized according to thepresent invention, in that an ester compound of the general formula##STR3## wherein: R₂, R₃, A and B have the above-defined meanings,

R₅ is a protected hydroxy group, and

R₆ is a (C₁ -C₆)alkyl group;

is reacted with an organometallic compound of the general formula

    R.sub.4 M(X).sub.p                                         (III)

wherein:

R₄ has the above meaning,

X is Cl, Br or I,

M is a metal selected from Li and Mg, and

p is, dependent on the valence of M, 0 or 1;

followed by deprotection.

In an equally attractive manner the C₁₇ -side chain can first befinalized. Therefore the invention also relates to a method of preparinga vitamin D compound as defined above, which method is characterizedaccording to the present invention, in that an ester compound of thegeneral formula ##STR4## wherein: R₂, R₃ and R₆ have the above meanings,and

R₅ ' is an optionally protected hydroxy group;

is reacted with an organometallic compound of the general formula

    R.sub.4 M(X).sub.p                                         (III)

wherein the symbols have the above meanings; after which the hydrindanecompound obtained, having the general formula ##STR5## is deprotected,if R₅ ' is a protected hydroxy group, and then oxidized to thecorresponding hydrindane-4-one compound of the general formula ##STR6##which compound of formula V, if desired after protection of the hydroxygroup, is then converted either

(a) with a Wittig reagent of the general formula ##STR7## wherein R₁ 'is a hydrogen atom or a protected hydroxy group, and the other symbolshave the above meanings;

or (b), after enolization and derivatization of the enolic hydroxygroup, with an enyne compound of the general formula ##STR8## whereinthe symbols have the above meanings, followed by hydrogenation andisomerization, to produce a compound of the general formula I, wherein Aand B form together a methylene group; followed by deprotection.

Hydroxy groups in the above intermediates or reactants may be protectedby a reaction with a suitable esterification or etherification agent. Asuitable esterification agent is an alkylchlorocarbonate having 2 to 5carbon atoms, or an aromatic carboxylic acid or saturated aliphaticcarboxylic acid having 1 to 4 carbon atoms such as benzoic acid, or aderivative of such acids suitable for the esterification reaction. Inorder to protect hydroxy groups in the form of an ether, in principleany etherification agent known for this purpose is suitable: forexample, a trialkylsilylimidazole, a trialkylsilylhalide, atrailkylsilyltriflate (-trifluoromethanesulfonate), adiphenylalkylsilylhalide, or a diphenylalkylsilyltriflate, or aderivative thereof, the alkyl groups of which have 1 to 6 carbon atoms.

Particularly suitable for this purpose are trimethylsilylchloride,tert.-butyldimethylsilylchloride,dimethyl-(1,1,2-trimethylpropyl)silylchloride, tert.-butyldimethylsilyltriflate, or trimethylsilylimidazole, because these etherificationagents readily react with the hydroxy group to be protected to form anether function, which on the one hand is sufficiently stable under theconditions of the reaction or reactions in view, but on the other handcan easily be removed [deprotection] to recover the original hydroxygroup; tert.-butyldimethylsilylchloride or triflate is to be preferred,because the tert.-butyldimethylsilyl group has been found to beexcellently suitable as a protective group.

The enolic hydroxy group is preferably derivatized by a reaction withN-phenyltriflimide to produce a triflate.

The starting compounds of formula II can conveniently be prepared fromreadily available substances, e.g. for the synthesis of vitamin Dcompounds with the above-defined preferred C₁₇ -side chains as follows:##STR9## wherein n is 1 or 2.

The introduction of a C₁₈ modification (R₂) into the vitamin D compoundof the invention can conveniently be achieved as described in theabove-mentioned EP-A-0521550.

Suitable examples of organometallic compounds of the above generalformula III are lithium compounds, such as isopropyllithium,cyclopropyllithium and tert.-butyllithium, and Grignard reagents, suchas isopropylmagnesium chloride, cyclopropylmagnesium chloride andtert.-butylmagnesium chloride, as well as the corresponding bromides.

The intermediate ester compound of the general formula IX, presentedabove, is new. Therefore the present invention also relates to thisintermediate, as well as to a method of preparing this compound. Theester compound of the general formula IX, wherein A and B form togethera methylene group, can conveniently be prepared by reacting an ester ofthe general formula ##STR10## wherein: R₂, R₃ and R₆ have the abovemeanings, and

R₇ is a derivatized hydroxy group,

is reacted with an enyne compounds of the general formula ##STR11##wherein the symbols have the above meanings; followed by hydrogenationand isomerization. This reaction is preferably carried out in tworeaction steps, viz. by first reacting the ingredients under theinfluence of an organic base such as triethylamine, and in the presenceof a palladium catalyst such as (PPh₃)₂ PdCl₂, and by then subjectingthe product obtained to a hydrogenation with hydrogen under theinfluence of a suitable catalyst such as Lindlar catalyst (Pd on CaCO₃,poisoned with lead), followed by an isomerization of the previtaminconfiguration obtained to the vitamin structure of the general formulaIX.

Alternatively, said ester compound of the general formula IX can easilybe synthetized by reacting a modified Windaus Grundmann ketone of thegeneral formula X or XI with a Wittig reagent as follows: ##STR12##wherein R₃ ═(CH₂)₃ or (CH₂)₄.

The symbols in the above formulas are defined hereinbefore. Theintermediate hydrindane-4-one compound of the general formula V,presented above, is new. Therefore the present invention also relates tothis intermediate and to a method of preparing this compound, viz. byoxidizing a hydrindane compound of the general formula IV, as definedabove, with an oxidizing agent, preferably selected from achromium-containing oxidant such as pyridinium chlorochromate orpyridinium dichromate, and ruthenium tetroxide.

The intermediate hydrindane compound of the general formula IV,presented above, is also new. Consequently, the present inventionrelates in addition to this intermediate and to a method of preparingthis compound, viz. by reacting a compound of the general formula II, asdefined above, with a metal-organic compound of the general formula III,as also defined above, in an inert organic solvent.

To improve the applicability of the new vitamin D compounds of theinvention for the above-described pharmacotherapeutic indications, thecompounds are usually processed to pharmaceutical compositions,comprising an effective amount of said vitamin D compound as the activeingredient in addition to a pharmaceutically acceptable carrier and/orat least one pharmaceutically acceptable auxiliary substance. Such acomposition may be delivered in a dosage unit form for oral, topical(dermal) or parenteral administration, comprising approx. 0.1 μg toapprox. 0.1 mg active ingredient per dosage unit. A composition fordiagnostic purposes may comprise, in addition to the vitamin D compoundof the present invention, a compatible, non-toxic carrier and/or atleast one auxiliary substance. A cosmetical composition may comprise, inaddition to an effective amount (in the range of approx. 0.1 μg toapprox. 0.1 mg per dosage unit in a dosage unit form) of the vitamin Dcompound of the present invention, a cosmetically acceptable, non-toxiccarrier and/or at least one auxiliary substance.

Finally the invention relates to a method for the treatment andprophylaxis of a number of disease states including autoiummune diseases(including diabetes mellitus), acne, alopecia, skin aging (includingphoto-aging), imbalance in the immune system, inflammatory diseases suchas rheumatoid arthritis and asthma, as well as diseases related toabnormal cell differentiation and/or proliferation, in a warm-bloodedliving being, comprising administering to said being or treating saidbeing with a pharmaceutical composition as defined above in a quantityeffective for the intended pupose. Examples of such diseases arepsoriasis and other hyperproliferative skin diseases.

The present invention also relates to the use of the abovepharmaceutical compositions for the treatment of solid, skin and bloodcancers, in particular of blood cancers such as leukemia, of breastcancer, and of skin cancers such as melanoma and squamous cellcarcinoma.

The above-defined cosmetical compositions, in particular selected fromthe group consisting of creams, lotions, ointments, liposomes and gels,can be used for the treatment and prevention of a number of skindisorders, such as inadequate skin firmness or texture, insufficientskin hydration, wrinkles and insufficient sebum secretion.

The invention will now be described in greater detail with reference tothe following specific Examples.

EXAMPLES Example I

Preparation of vitamin ester ##STR13##

(a). Ph₃ P (6.5 g) and imidazole (4.8 g) are added to a solution of diol(1) (5.0 g) in THF (100 ml). The suspension is cooled to -20° C., and I₂(6.28 g) is added in portions. After being stirred for 15 min, thereaction mixture is warmed to room temp., further stirred for 15 min.,cooled to 0° C., and poured into saturated aqueous NaHCO₃ (50 ml). Themixture is extracted with Et₂ O and the extract is washed with saturatedaqueous Na₂ S₂ O₃ and H₂ O, dried and filtered. Concentration affords aresidue which is purified by flash chromatography (8% EtOAc/hexane) togive 7.32 g of iodide (2). After crystallization from EtOAc/hexane, theproduct has a melting point of 51° C.; identification by NMR and elem.analysis.

(b). Pyridinium dichromate (8.66 g) is added to a solution of compound(2) (3.99 g) in 50 ml CH₂ Cl₂. The mixture is stirred for 6 h at roomtemp. Et₂ O (60 ml) is added, and the resulting suspension is stirredfor 15 min and filtered. The filtrate is washed with brine, dried,filtered and concentrated. Purification by flash chromatography (10%EtOAc/hexane) gives the desired iodo ketone (3) in a yield of 3.57 g;crystallization from Et₂ O/hexane: m.p. 65° C. Identification by NMR andelem. analysis.

(c). Lithium diisopropylamine is prepared by addition of i-Pr₂ NH (3.9mmol) to a cooled (-78° C.) solution of n-BuLi in hexane (3.5 mmol in1.43 ml). After stirring for 10 min, the mixture is diluted with THF (4ml), stirred at 0° C. for 30 min, and cooled to -78° C. A solution ofketone (3) (1.0 g) in 14 ml THF is slowly added, followed by a solutionof N-phenyltriflimide (1.225 g) in 4 ml THF. The mixture is stirred for2 h at -78° C. After being warmed to 0° C., the reaction is quenched byaddition of a few drops of MeOH and water. Concentration gives a crudeproduct which is diluted with EtOAc/hexane (30 ml), washed with brine,dried, filtered and concentrated. The resulting residue is purified byflash chromatography (2% EtOAc/hexane), affording 1.29 g of the iodotriflate (4) as a colourless oil. Identification by NMR and elem.analysis.

(d). A suspension of CuI (201 mg) and Zn (161 mg) in EtOH/H₂ O (6 ml7:3; deoxygenated) is sonicated for 5 min. Methyl acrylate (637 μl,freshly distilled), and a solution of the iodide (4) (160 mg) in EtOH/H₂O (1 ml 7:3) are successively added, and the resulting mixture issonicated for 40 min. Dilution with Et₂ O (15 ml) and filtration gives asolution that is washed with brine. The aqueous phase is extracted withEt₂ O (30 ml) and the combined organic extracts are dried, filtered andconcentrated. Flash chromatography of the residue (6% EtOAc/hexane)affords 96 mg of the methyl ester (5) (colourless oil). Identificationof NMR and elem. analysis.

(e). The palladium-catalyzed coupling between vinyl triflate (5) and theenyne (6) is performed as follows: A mixture of enyne (6) (507 mg),triflate (5) (500 mg), Et₃ N (4.85 mmol) and (Ph₃ P)₂ PdCl₂ (16 mg) in21 ml DMF is heated at 75° C. for 1 h. The mixture is cooled to roomtemp., diluted with EtOAc/hexane (50 ml 1:3), and washed with brine.Drying, filtration and concentration gives a residue which is purifiedby flash chromatography (2-4% Et₂ O/hexane) to afford 675 mg of dienyne(7) (viscous liquid). Identification by NMR.

(f). Product (7) is hydrogenated by Lindlar catalyst as follows: Asolution (0.2 ml) of 50 μl quinoline in 10 ml hexane is added to asolution of dienyne (7) (305 mg) in 12 ml hexane. Lindlar catalyst (50mg), previously dried, is added and the resulting solution is exposed tohydrogen gas at atmospheric pressure. After stirring for 8 h, thereaction mixture is filtered and concentrated. The residue is purifiedby flash chromatography (1-3% Et₂ O/hexane) to give 295 mg of theprotected previtamin D compound. Isomerization of previtamin D compoundto vitamin D compound (8): The previtamin D compound obtained (295 mg )is dissolved in 15 ml isooctane and refluxed in the dark for 5 h.Concentration gives a residue which is purified by flash chromatography(2-4% Et₂ O/hexane) to afford 290 mg of compound (8). the product isidentified by ¹ H-NMR, ¹³ C-NMR and elem. analysis.

¹ H-NMR (δ, CDCl₃): 6.24 and 6.02 (d, 2H), 5.18 (m, 1H), 4.87 (m, 1H),4.37 (m, 1H), 4.18 (m, 1H), 3.67 (s, 3H), 0.93 (d, 3H), 0.88 (s, 18H),0.53 (s, 3H), 0.07 (s, 12H). ¹³ C-NMR (δ, CDCl₃): 173.1, 148.4, 141.0,135.0, 132.2, 118.0, 111.2, 72.1, 67.5, 56.3, 51.3, 46.0, 45.7, 44.8,40.6, 35.8, 35.3, 34.4, 31.5, 28.8, 27.6, 25.8, 25.7, 23.4, 22.6, 22.1,21.5, 18.7, 18.1, 18.0, 14.0, 11.9, -4.8, -4.8, -4.9, -5.2. Elem. anal.:Calcd. for C₃₈ H₆₆ O₄ Si₂ : C, 70,75; H, 10.62. Found: C, 70.42; H,10.43.

In a corresponding manner the following ester compounds are prepared:

    ______________________________________                                        general formula                                                                ##STR14##                                                                    compound no.   R.sub.2  R.sub.3    R.sub.6                                    ______________________________________                                         (9)           CH.sub.3 (CH.sub.2).sub.4                                                                         C.sub.2 H.sub.5                            (10)           C.sub.2 H.sub.5                                                                        (CH.sub.2).sub.3                                                                         C.sub.2 H.sub.5                            ______________________________________                                    

Compd. (9): This 24-homo compound is prepared by using in the above step(a) as the starting substance a homologue of compd. (1), having a1-methyl-3-hydroxypropyl side chain; in step (d) ethyl acrylate is usedas the olefin.

¹ H-NMR (δ, CDCl₃): 0.08 (s, 12H), 0.52 (s, 3H), 0.87 (s, 18H), 0.90 (d,3H), 1.25 (t, 3H), 4.11 (q, 2H), 4.20 (m, 1H), 4.37 (m, 1H), 4.87 (d,1H), 5.18 (d, 1H), 6.01 (d, 1H), 6.24 (d, 1H).

Compd. (10): This 18-homo compound is prepared by using in the abovestep (a) as the starting compound a homologue of compd. (1), prepared asdescribed in published European patent application 521550 [compound no.(64)].

¹ H-NMR (δ, CDCl₃): 0.87 (s, 6H), 0.88 (t, 3H), 1.01 (d, 3H), 1.25 (t,3H), 1.98 (t, 1H), 2.25 (m, 2H), 2.44 (dd, 1H), 4.13 (q, 2H), 4.18 (m,1H), 4.37 (m, 1H), 4.86 (s, 1H), 5.17 (s, 1H), 6.01 (d, 1H), 6.23 (d,1H).

Example II

Preparation of vitamin D compound from vitamin ester ##STR15##

Compound (11) is prepared as follows: Cyclopropyl bromide (0.51 mmol) isslowly added to a cooled (-20° C.) solution of t-BuLi in Et₂ O (0.51mmol in 0.602 ml), to produce cyclopropyllithium. The resulting mixtureis warmed to room temp. and diluted with 2.4 ml Et₂ O. 1 ml of thissolution is slowly added to a cooled (-78° C.) solution of compd. (8)(50 mg) in 3 ml Et₂ O. The reaction mixture is allowed to come to -40°C. and quenched with a few drops of water. The resulting solution isdiluted with Et₂ O, washed with brine, dried, filtered and concentrated.The concentrate is filtered through a flash chromatography column (2%Et₂ O/hexane), affording a product (46 mg) which is dissolved in 7 mlTHF and stirred in the dark at room temp. with tetrabutylammoniumfluoride in THF (0.36 mmol in 0.36 ml) for 24 h. concentrationgives a residue which is diluted with EtOAc (20 ml), dried, filtered,concentrated and flash chromatographed (60% EtOAc/hexane) to give 23 mgof the desired compound (11) as a white solid.

¹ H-NMR (δ, CD₂ Cl₂): 6.44 and 5.99 (d, 2H), 5.27 (br-d, 1H), 4.95(br-d, 1H), 4.35 (m, 1H), 4.15 (m, 1H), 0.92 (d, 3H), 0.81 (m, 2H), 0.53(s, 3H), 0.34 (m, 8H). ¹³ C-NMR (δ, CD₂ Cl₂): 148.6, 143.5, 133.9,125.1, 117.6, 111.8, 71.2, 71.0, 67.2, 57.2, 56.8, 45.8, 43.5, 43.4,41.0, 37.1, 36.6, 29.4, 28.0, 24.0, 22.6, 20.8, 19.0, 12.1, 0.8, -0.5.

In a corresponding manner the following vitamin D compounds areprepared:

    ______________________________________                                        general formula                                                                ##STR16##                                                                    compound no. R.sub.2   R.sub.3   R.sub.4                                      ______________________________________                                        (12)         CH.sub.3  (CH.sub.2).sub.3                                                                        C(CH.sub.3).sub.3                            (13)         CH.sub.3  (CH.sub.2).sub.3                                                                        CH(CH.sub.3).sub.2                           (14)         C.sub.2 H.sub.5                                                                         (CH.sub.2).sub.3                                                                        CH(CH.sub.3).sub.2                           (15)         C.sub.2 H.sub.5                                                                         (CH.sub.2).sub.3                                                                        cyclopropyl                                  (16)         CH.sub.3  (CH.sub.2).sub.4                                                                        cyclopropyl                                  (17)         CH.sub.3  (CH.sub.2).sub.4                                                                        CH(CH.sub.3).sub.2                           ______________________________________                                    

Compd. (12) is prepared by using t-butyllithium instead ofcyclopropyllithium.

Compounds (13), (14) and (17) are prepared by using isopropyllithiuminstead of cyclopropyllithium.

Compd. (12): ¹ H-NMR (δ, CDCl₃): 6.38 and 6.01 (d, 2H), 5.33 (m, 1H),5.00 (m, 1H), 4.43 (m, 1H), 4.23 (m, 1H), 1.00 (s, 18H), 0.93 (d, 3H),0.54 (s, 3H). ¹³ C-NMR (δ, CDCl₃): 148.7, 143.4, 134.0, 125.0, 117.6,111.8, 80.0, 71.1, 67.1, 60.6, 56.9, 56.8, 46.3, 45.8, 43.4, 42.8, 40.9,37.0, 36.5, 34.2, 29.4, 28.8, 28.0, 24.0, 23.3, 22.7, 19.2, 12.1.

Compd. (13): ¹ H-NMR (δ, CD₃ OD): 0.55 (s, 3H), 0.90 (m, 15H), 2.22 (dd,1H), 2.48 (dd, 1H), 2.83 (dd, 1H), 4.09 (m, 1H), 4.31 (t, 1H), 4.86 (b?,1H), 5.25 (b, 1H), 6.05 (d, 1H), 6.29 (d, 1H).

Compd. (14): ¹ H-NMR (δ, CDCl₃): 0.84 (t, 3H), 0.95 (m, 12H), 1.00 (d,3H), 2.32 (dd, 1H), 2.60 (dd, 1H), 2.83 (dd, 1H), 4.24 (m, 1H), 4.44 (m,1H), 5.01 (b, 1H), 5.33 (b, 1H), 6.01 (d, 1H), 6.39 (d, 1H).

Compd. (15): ¹ H-NMR (δ, CD₃ OD): 0.70-0.40 (m, 8H), 0.76 (m, 2H), 0.91(t, 3H), 1.01 (d, 3H), 2.00 (m, 1H), 2.22 (dd, 1H), 2.29 (b, 1H), 2.48(dd, 1H), 2.83 (dd, 1H), 4.09 (m, 1H), 4.31 (t, 1H), 4.85 (b, 1H), 5.25(b, 1H), 6.04 (d, 1H), 6.29 (d, 1H).

Compd. (16): ¹ H-NMR (δ, CD₃ OD): 0.20-0.40 (m, 8H), 0.57 (s, 3H), 0.79(m, 2H), 0.95 (d, 3H), 2.25 (dd, 1H), 2.51 (dd, 1H), 2.86 (dd, 1H), 4.12(m, 1H), 4.35 (t, 1H), 4.89 (b, 1H), 5.28 (b, 1H), 6.08 (d, 1H), 6.32(d, 1H).

Compd. (17): ¹ H-NMR (δ, CD₃ OD): 0.53 (s, 3H), 0.90 (m, 12H), 2.22 (dd,1H), 2.48 (dd, 1H), 2.83 (dd, 1H), 4.09 (m, 1H), 4.31 (t, 1H), 4.86 (b,1H), 5.25 (b, 1H), 6.05 (d, 1H), 6.29 (d, 1H).

Example III

Preparation of1-(1-methyl-5-hydroxy-5,5-diisopropylpentyl)-hydrindanol-4 (19)##STR17##

(a). Starting compound (1) is converted to the corresponding iodide (2)as described in Example I(a).

(b). The compound (2) obtained is converted in a corresponding manner asdescribed in Example I(d), using ethyl acrylate as the olefin, toproduce ester compound (18).

(c). Ester compound (18) is converted to compound (19) by a reactionwith an excess of isopropyllithium, in a corresponding manner asdescribed in Example II. The product is identified by ¹ H-NMR.

In a corresponding manner the following compounds are prepared:

    ______________________________________                                        general formula                                                                ##STR18##                                                                    compound no. R.sub.2   R.sub.3   R.sub.4                                      ______________________________________                                        (20)         CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                        cyclopropyl                                  (21)         CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                        isopropyl                                    (22)         CH.sub.3  (CH.sub.2).sub.4                                                                        isopropyl                                    (23)         CH.sub.3  (CH.sub.2).sub.4                                                                        cyclopropyl                                  ______________________________________                                    

The products are identified by ¹ H-NMR.

Example IV

Preparation of1-(1-methyl-5-hydroxy-5,5-diisopropylpentyl)-hydrindanone-4 (24)##STR19##

Oxidation of compound (19) by using pyridinium dichloromate as theoxidant, in a corresponding manner as described in Example I(b), affordsthe desired ketone (24) in a yield of 84%. The product is identified by¹ H-NMR.

In a corresponding manner the following ketones are prepared:

    ______________________________________                                        general formula                                                                ##STR20##                                                                    compound no. R.sub.2   R.sub.3   R.sub.4                                      ______________________________________                                        (25)         CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                        cyclopropyl                                  (26)         CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                        isopropyl                                    (27)         CH.sub.3  (CH.sub.2).sub.4                                                                        isopropyl                                    (28)         CH.sub.3  (CH.sub.2).sub.4                                                                        cyclopropyl                                  ______________________________________                                    

The products are identified by ¹ H-NMR.

Example V

Preparation of vitamin D compound (13) from ketone (24) ##STR21##

(a). The free hydroxy group is protected by a reaction withtrimethylsilyltriflate (TBS-triflate) in the presence of triethylamineand in methylene chloride as the solvent; temp. -78° C.--0° C.; yield ofcompound (29) is 80%.

(b) The enolisation is carried out in a corresponding manner asdescribed in Example I(c), producing compound (30) in a yield of 71%.

(c). In a corresponding manner as described in Example I(e), thecoupling reaction with enyne (6) is performed, affording compound (31)in a yield of 94%.

(d). The final reaction step is carried out in a corresponding manner asdescribed in Example I(f), followed by deprotection (desilylation), asdescribed in Example II, with tetrabutyl ammoniumfluoride. The finalvitamin D compound (13) is obtained in an overall yield of 65%. Theproduct is identical with the product obtained according to Example II.

In a corresponding manner the following vitamin D compounds areprepared.

    ______________________________________                                        general formula                                                                ##STR22##                                                                    compound no. R.sub.2   R.sub.3   R.sub.4                                      ______________________________________                                        (32)         CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                        cyclopropyl                                  (33)         CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                        isopropyl                                    (17)         CH.sub.3  (CH.sub.2).sub.4                                                                        isopropyl                                    (16)         CH.sub.3  (CH.sub.2).sub.4                                                                        cyclopropyl                                  ______________________________________                                    

The products are identified by ¹ H-NMR. The last two vitamin D compoundsare identical with the corresponding vitamin D compounds preparedaccording to Example II.

Example VI

Preparation of 19-nor-vitamin D compound (35) from ketone (29) ##STR23##A solution of 1.14 g (2 mmol) of phosphine oxide (34) in 15 ml of dryTHF is cooled to -78° C. n-Buthyllithium (BuLi), as a 2.5M solution inhexane, is added dropwise until the red colour persists. Then 0.8 ml ofa 2.5M solution of BuLi is added. Stirring is continued for 15 min,followed by the dropwise addition of 0.73 g (1.8 mmol) of ketone (29) in5 ml THF. After another hour of stirring, the reaction mixture allowedto reach 0° C. and then quenched by the addition of 50 ml of a saturatedNH₄ Cl-solution. Extractive work-up and flash chromatography (2% EtOAcin hexane) then affords the protected diene compound. Desilylation byreaction with 10 eq. of tetrabutylammonium fluoride (TBAF.3aq) in THF(10 ml) during 48 hours gives compound (35), which is purified by flashchromatography using EtOAc as an eluent, followed by recrystallizationfrom MeOH/EtOAc. The overall yield is 53%. Identification by ¹ H-NMR.

In a corresponding manner the following vitamin D compounds areprepared:

    ______________________________________                                        general formula                                                                ##STR24##                                                                    compound no.                                                                           R.sub.2   R.sub.3  R.sub.4  A    B                                   ______________________________________                                        (36)     CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                       isopropyl                                                                              H    H                                   (37)     CH═CH.sub.2                                                                         (CH.sub.2).sub.3                                                                       cyclopropyl                                                                            H    H                                   (38)     CH.sub.2 OH                                                                             (CH.sub.2).sub.3                                                                       isopropyl                                                                              H    H                                   ______________________________________                                    

The products are identified by ¹ H-NMR.

Example VII

Affinity to intracellular vitamin D receptor

Vitamin D compounds according to the invention are dissolved in ethanolin concentrations ranging from 10⁻¹³ to 10⁻⁷ M. The affinity towards thecalf thymus intracellular vitamin D receptor (VDR) is determined in abiological assay. In this assay, ³ H-1α,25-dihydroxycholecalciferol (³H-1α,25-DHCC), which is specifically bound to the VDR, is replaced bythe tested compounds. Especially the tested compounds 11, 13 and 14 havea very high VDR-affinity. A high VDR-affinity is indicative forbiologically active substances.

Example VIII

Affinity to vitamin D binding protein

Vitamin D binding protein (DBP) is the specific carrier for vitamin Dand its metabolites in blood. The biological activity of vitamin Dcompounds depends on their binding to DBP, because strong binding to DBPwill reduce the intracellular access to the VDR. Binding to the DBP mayalso influence the half-life of the vitamin D derivatives incirculation. Weak binders are rapidly matabolized, which is a favourableaspect in topical application. In the assay DBP is incubated with ³H-1α,25-DHCC and 1α,25-DHCC or with several vitamin D compoundsaccording to the invention. To this purpose, the vitamin compounds aredissolved in ethanol in concentrations ranging from 10⁻¹¹ to 2.5×10⁻⁶ M.The percentage bound/unbound ³ H-1α,25-DHCC is then calculated. DBP ispurified from total human serum. The results are shown in the appendedFIGS. 1 and 2. FIGS. 1 and 2 show the binding of vitamin D compounds tohuman vitamin D binding protein. [³ H]1α,25(OH)₂ D₃ =³ H-1α,25-DHCC; inboth Figures =1α,25-DHCC (known compound); in FIG. 1 ▴=compound 13 and∇=compound 14; in FIG. 2 Δ=compound 11. Compounds 14 and 11 bind ratherweakly to the DBP, compared to the known 1α,25-DHCC. Compound 13 is avery weak binder.

Example IX

Cell differentiation

Vitamin D compounds according to the invention are dissolved in ethanolin concentrations ranging from 10⁻¹² to 10⁻⁶ M and tested for theircapacity to induce cell differentiation in a HL-60 assay. In this assay,morphologic and biochemical examination of the human leukemic cell lineHL-60 is done, in order to establish whether cell differentiation hastaken place.

Differentiation is expressed as the maturation parameters nitrobluetetrazolium (NBT) reduction, non-specific esterase, and as thepercentage of mature cells beyond the myelocyte stage which is visibleafter staining with May-Grunwald Giemsa. After culturing with the known1α,25-DHCC or with vitamin D compounds of the invention, the percentageof cells containing black formazan deposits is determined. An increasein the percentage of NBT reducing cells indicates an increase in celldifferentiation.

Proliferation and vitality of the cell cultures are established bycounting the number of cells and by the trypan blue exclusion method.The vitality and proliferation of the cells in the HL-60 cultures aregood in all conditions tested. 1α,25-DHCC (known), compound 12, compound11, compound 13 and compound 14 all induce differentiation andmaturation of the HL-60 cells. In the cytological test (non-specificesterase and May-Grunwald Giemsa) especially compounds 13 and 11 aregood differentiators. The optimum effect is found at concentrations inthe range of 10⁻⁸ to 10⁻⁷ M. The NBT-reduction inducing capacity ofcompounds 13 and 14 is about 10 × stronger than that of the known1α,25-DHCC. Compounds 11 and 12 are about 5× more potent in inducingNBT-reduction than 1α,25-DHCC (FIGS. 3 and 4).

The above implies that the tested new vitamin D compounds of theinvention display a higher cell differentiating activity than the known1α,25-DHCC. FIGS. 3 and 4 (appended) show the differentiating effect ofthe tested vitamin D compounds on human leukemia cells of the HL-60line. In both Figures =1α,25-DHCC; in FIG. 3 ◯ is compound 13 and ∇ iscompound 14; in FIG. 4 Δ=compound 11 and ∇=compound 12.

Example X

Calciotropic effect

The most well-known effect of 1α,25-DHCC is its action on the calciummetabolism, the calciotropic effect. Calciotropic target organs are theintestine, the bone and the kidney. The vitamin D compounds according tothe invention are dissolved in ethanol and tested in the so-calledCaco-2 assay for intestinal calcium (Ca) transport. In this assay, thevitamin D-induced influx of ⁴⁵ Ca²⁺ is measured in monolayers of theintestinal cancer cell line Caco-2. This influx is corrected for theconcentration-driven Ca²⁺ influx and is a measure for the Ca transportacross the intestinal wall. The Caco-2 cells are known to have vitamin Dreceptors. Increased intestinal calcium transport can be the first stepleading to a rise in blood calcium levels (and eventually tohypercalcemia). In Table A below the effects of vitamin D compounds ofthe invention, compared with the known 1α,25-DHCC, on the Ca²⁺ influx inintestinal Caco-2 cell cultures are presented. The values in the tablerepresent the relative increase in Ca² + influx (the value for1α,25-DHCC is arbitrarily fixed at 100). The results in Table Ademonstrate that compound 11, compound 13 and compound 14 are weakerstimulators of intestinal calcium absorption than 1α,25-DHCC.

                  TABLE A                                                         ______________________________________                                               compound 10.sup.-9 M                                                   ______________________________________                                               experiment 1:                                                                 1α,25-DHCC                                                                       100                                                                  compound 11                                                                            79                                                                   compound 12                                                                            99                                                                   experiment 2:                                                                 1α,25-DHCC                                                                       100                                                                  compound 13                                                                            65                                                                   compound 14                                                                            78                                                            ______________________________________                                    

Example XI

Calciotropic effect

Together with the intestine and the bone, the kidney is one of the majortarget organs of 1α,25-DHCC. The kidney plays an extremely importantrole in calcium homeostasis, since about 98% of the calcium has to bereabsorbed in the kidneys in order to prevent calcium loss andhypocalcemia. The vitamin D compounds according to the invention aredissolved in ethanol and tested in the rabbit kidney cell assay. In thisassay, reabsorption of ⁴⁵ Ca²⁺ is measured in monolayers of rabbitkidney cells. The cells are isolated by immunodissection of connectingtubules with the aid of monoclonal antibodies. In Table B below theeffects of vitamin D compounds of the invention, compared with the known1α,25-DHCC, on the Ca²⁺ -reabsorption in rabbit renal cell cultures arepresented. The values in the Table represent the increase in Ca²⁺-reabsorption in mmol/cm² /h. The results in Table B demonstrate thatcompound 13 and compound 11 are weaker stimulators of renalCa-reabsorption than compound 12 and 1α,25-DHCC itself.

                  TABLE B                                                         ______________________________________                                               compound 10.sup.-9 M                                                   ______________________________________                                               1α,25-DHCC                                                                       13.4                                                                 compound 11                                                                            2.4                                                                  compound 12                                                                            12.8                                                                 compound 13                                                                            -3.7                                                          ______________________________________                                    

Example XII

Cell differentiation versus calciotropic effect

One of the set-backs of the highly active vitamin D compounds, such asthe well-known 1α,25-DHCC, is its calciotropic effect, which may lead totoxic hypercalciuria, hypercalcemia and urolithiasis. Therefore, itshould be very advantageous to develop compounds with a high selectivityof biological action. In other words, compounds in which the ratiobetween the induction of cell differentiation and calciotropic effects,e.g. the stimulation of intestinal Ca transport, is changed compared to1α,25-DHCC. The ratio between differentiation-inducing capacity and thestimulation of intestinal Ca transport is defined as the fractionbetween the concentration at which 50% NBT reduction in HL-60 cells isobtained, and the concentration at which a half-maximal increase inintestinal Ca²⁺ transport is reached. The smaller the ratio, the higherthe relative cell differentiating capacity. The results are presented inTable C.

                  TABLE C                                                         ______________________________________                                        compound       fraction                                                                              ratio relative to DHCC                                 ______________________________________                                        1α,25-DHCC                                                                             107     1.00                                                   compound 12    29.7    0.28                                                   compound 11    14.4    0.13                                                   compound 14    2.6     0.02                                                   compound 13    2.5     0.02                                                   ______________________________________                                    

Table C shows, that the tested new vitamin D compounds of the inventionhave better relative cell differentiating properties than the known1α,25-DHCC. Compound 14 and compound 13 have 50× more selective actionsthan 1α,25-DHCC. Compound 11 has a 8× better ratio. Compound 12 has a 4×better ratio. This makes the new vitamin D compounds of the presentinvention extremely suitable for applications where the differentiationof cells (such as in hyperproliferative conditions) is desired.

We claim:
 1. A method for the treatment and prophylaxis of diseasestates selected from the group consisting of autoimmune diseases,.[.ache, alopecia,.]. .Iadd.acne, .Iaddend.skin aging, imbalance in theimmune system, inflammatory diseases and diseases related to abnormalcell differentiation or proliferation, in a warm-blooded living being,comprising administering to said being or treating said being with acomposition comprising at least one member selected from the groupconsisting of a pharmaceutically acceptable carrier and pharmaceuticallyacceptable auxiliary substances and mixtures thereof, and, as an activeingredient a compound of formula (1) ##STR25## wherein R₁ is a hydrogenatom or a hydroxy group;R₂ is a (C₁ -C₃)alkyl group, a hydroxy(C₁-C₃)alkyl group, a (C₁ -C₂)alkoxymethyl group or a (C₂ -C₃)alkenyl oralkynyl group; R₃ is a branched or non-branched, saturated orunsaturated aliphatic 3- to 5-membered hydrocarbon or oxahydrocarbonbiradical, having at least 3 atoms in the main chain and beingoptionally substituted with one or more substituents selected fromepoxy, fluoro and hydroxyl R₄ is a sec. or tert. (C₃ -C₆)alkyl group ora (C₃ -C₆)cycloalkyl group; and A and B are each individually hydrogenatoms or .[.ethyl.]. .Iadd.methyl .Iaddend.groups, or A and B formtogether a methylene group in a quantity effective for the intendedpurpose.
 2. A method as claimed in claim 1 wherein the disease treatedis psoriasis which is a disease of abnormal cell proliferation.
 3. Amethod of claim 2 wherein the active ingredient is of the formula##STR26##
 4. A method for the treatment and prevention of skin disordersin a warm-blooded living being, comprising treating said being with acosmetical composition comprising at least one member selected from thegroup consisting of conventionally acceptable, non-toxic carriers andauxiliary substances and mixtures thereof, and, as the active ingredienta compound of formula (I) wherein:R₁ is a hydrogen atom or a hydroxygroup; R₂ is a (C₄ -C₃)alkyl group, a hydroxy(C₁ -C₃)alkyl group, a (C₁-C₂)alkoxymethyl group or a (C₂ -C₃)alkenyl or alkynyl group; R₃ is abranched or non-branched, saturated or unsaturated aliphatic 3- to5-membered hydrocarbon or oxahydrocarbon biradical, having at least 3atoms in the main chain and being optionally substituted with one ormore substituents selected from epoxy, fluoro and hydroxy; R₄ is a sec.or tert. (C₃ -C₆)alkyl group or a (C₃ -C₆)cycloalkyl group; and A and Bare each individually hydrogen atoms or .[.ethyl.]. .Iadd.methyl.Iaddend.groups, or A and B form together a methylene group in aneffective quantity.
 5. A method for the treatment and prophylaxis ofdisease states selected from the group consisting of autoimmunediseases, .[.ache, alopecia,.]. .Iadd.acne, .Iaddend.skin aging,imbalance in the immune system, inflammatory diseases and diseasesrelated to abnormal cell differentiation or proliferation, in awarm-blooded living being, comprising administering to said being ortreating said being with a composition comprising at least one memberselected from the group consisting of a pharmaceutically acceptablecarrier and pharmaceutically acceptable auxiliary substances andmixtures thereof, and, as an active ingredient a compound of formula (1)##STR27## wherein: R₁ is a hydroxy group;R₂ is a (C₁ -C₃)alkyl group, ahydroxy(C₁ -C₃)alkyl group, a (C₁ -C₂)alkoxymethyl group or a (C₂-C₃)alkenyl or alkynyl group; R₃ is a biradical of the formula

    --O--CH.sub.2 --(CH.sub.2).sub.n --, --CH.sub.2 --CH.sub.2 --(CH.sub.2).sub.n --, --CH═CH--(CH.sub.2).sub.n -- or --CH.sub.2 --CH.sub.2 --CH(CH.sub.3)--,

wherein n is 1 or 2; R₄ is an isopropyl group, a cyclopropyl group or atert.-butyl group; and A and B are hydrogen atoms or form together amethylene group in a quantity effective for the intended purpose.
 6. Amethod for the treatment and prophylaxis of disease states selected fromthe group consisting of autoimmune diseases, acne, .[.alopecia,.]. skinaging, imbalance in the immune system, inflammatory diseases anddiseases related to abnormal cell differentiation or proliferation, in awarm-blooded living being, comprising administering to said being ortreating said being with a composition comprising at least one memberselected from the group consisting of a pharmaceutically acceptablecarrier and pharmaceutically acceptable auxiliary substances andmixtures thereof, and, as an active ingredient at least one compound offormula (1) ##STR28## wherein: R₁ is a hydrogen atom or a hydroxygroup;R₃ is a branched or non-branched, saturated or unsaturatedaliphatic 3- to 5-branched hydrocarbon or oxahydrocarbon biradical,having at least 3 atoms in the main chain and being optionallysubstituted with one or more substituents selected from epoxy, fluoroand hydroxy; R₄ is an isopropyl group A and B are each individuallyhydrogen atoms or .[.ethyl.]. .Iadd.methyl .Iaddend.groups, or A and Bform together a methylene group in a quantity effective for the intendedpurpose.
 7. A method as claimed in claim 5 wherein the disease treatedis psoriasis which is a disease of abnormal cell proliferation.
 8. Amethod as claimed in claim 6 wherein the disease treated is psoriasiswhich is a disease of abnormal cell proliferation.
 9. A method for thetreatment and prevention of skin disorders in a warm-blooded livingbeing, comprising treating said being with a cosmetical compositioncomprising at least one member selected from the group consisting ofconventionally acceptable, non-toxic carriers and auxiliary substancesand mixtures thereof, and, as the active ingredient a compound offormula (I) ##STR29## wherein: R₁ is a hydroxy group;R₂ is a (C₁-C₃)alkyl group, a hydroxy(C₁ -C₃)alkyl group, a (C₁ -C₂)alkoxymethylgroup or a (C₂ -C₃)alkenyl or alkynyl group; R₃ is a biradical of theformula

    --O--CH.sub.2 --(CH.sub.2).sub.n --, --CH.sub.2 --C.sub.2 --(CH.sub.2).sub.n --, --CH═CH--(CH.sub.2).sub.n -- or --CH.sub.2 --CH.sub.2 --CH(CH.sub.3)--,

wherein n is 1 or 2; R₄ is an isopropyl group, a cyclopropyl group or atert.-butyl group; and A and B are hydrogen atoms or form together amethylene group in a quantity effective for the intended purpose.
 10. Amethod for the treatment and prevention of skin disorders in awarm-blooded living being, comprising treating said being with acosmetical composition comprising at least one member selected from thegroup consisting of conventionally acceptable, non-toxic carriers andauxiliary substances and mixtures thereof, and, as the active ingredienta compound of formula (I) ##STR30## wherein: R₁ is a hydrogen atom or ahydroxy group;R₃ is a branched or non-branched, saturated or unsaturatedaliphatic 3- to 5-membered hydrocarbon or oxahydrocarbon biradical,having at least 3 atoms in the main chain and being optionallysubstituted with one or more substituents selected from epoxy, fluoroand hydroxy; R₄ is an isopropyl group A and B are each individuallyhydrogen atoms or ethyl groups, or A and B form together a methylenegroup in a quantity effective for the intended purpose.
 11. A method ofclaim 5 wherein the active ingredient is of the formula ##STR31##
 12. Amethod of claim 6 wherein the active ingredient is of the formula